English
聯系我們
網站地圖
郵箱
舊版回顧



科學家破解基孔肯雅病毒入侵機制

文章來源:微生物研究所   發布時間:2019-05-10  【字號:     】  

  59日,中國科學院高福團隊在《細胞》(Cell雜志上發表了題為Molecular Basis of Arthritogenic Alphavirus Receptor MXRA8 Binding to Chikungunya Virus Envelope Protein 的文章,首次從分子水平闡釋了基孔肯雅病毒囊膜表面E蛋白與其細胞受體MXRA8分子的相互作用機制,揭示了此類病毒入侵細胞的分子機制,為抗病毒藥物開發及新型疫苗設計提供了新靶點。

  基孔肯雅熱(Chikungunya fever, CHIKF)是一種由基孔肯雅病毒(Chikungunya virusCHIKV)引起,經蚊蟲叮咬傳播的急性發熱性傳染病。該病毒病過去主要分布在非洲、南亞、東南亞熱帶和亞熱帶地區。近年來,隨著全球變暖和現代交通工具等因素,該病毒疫情呈不斷暴發和蔓延的趨勢。2005年以來疫情在東南亞和南亞地區再度暴發,2007年疫情傳播至歐洲,2013年疫情開始在加勒比海地區、美國以及南美洲等地暴發流行,造成了數百萬人感染。人感染該病毒后可致急性或慢性的外周關節痛或關節炎,嚴重時致人死亡,目前尚無特異性的治療方法和疫苗,使得該病毒引起人們的廣泛關注,也是重要的全球性公共衛生問題之一。與該病毒親緣關系較近,同樣能引起關節炎的甲病毒還包括馬亞羅病毒(Mayaro virus)、羅斯河病毒(Ross River virus)、阿尼昂尼昂病毒(O' nyong- nyong virus)等。

  病毒侵入細胞的第一步依賴于病毒囊膜蛋白與宿主受體的相互作用。過去的研究認為,CHIKV等多種致關節炎甲病毒囊膜表面含有80個三聚體形式的刺突E蛋白,每個三聚體由三個E1/E2二聚體組成其中E1蛋白參與膜融合,E2蛋白參與受體的結合。近來研究報道表明,基質重塑相關蛋白8Matrix remodeling-associated protein 8MXRA8)分子是CHIKV等多種致關節炎甲病毒的受體。MXRA8是一種在上皮細胞、髓細胞及間充質干細胞表達的黏附分子,廣泛分布于形成軟骨、肌肉和骨骼的細胞表面。但是,MXRA8分子如何介導病毒入侵卻是未解之謎。

  高福團隊深入研究了MXRA8分子介導CHIKV的入侵機制。首先,他們利用結構生物學的技術和方法解析了小鼠MXRA8mMXRA8)的晶體結構,表明MXRA8分子胞外段由兩個免疫球蛋白(Ig)樣結構域構成,但其兩個Ig樣結構域呈現獨特的拓撲結構。結構域1D1)由兩個不連續的片段構成,而結構域2D2)插在了D1的兩個片段之間,這導致D1D2之間有兩個柔性鉸鏈(Hinge loop)連接,這與以往所有報道的含有兩個Ig樣結構域的蛋白都不同。因此,他們首次提出MXRA8是一種新型的Ig樣受體分子,并具有獨特的拓撲結構及結構域間組裝形式。

  為進一步闡明CHIKV E與受體MXRA8相互作用機制,研究團隊解析了人MXRA8hMXRA8)與CHIKV E復合物的晶體結構,他們發現MXRA8CHIKV E采用一種獨特的3:3的結合模式,MXRA8結合到病毒表面三聚體刺突蛋白兩個E蛋白單體間的“峽谷”中,形成非常緊密的結合模式。其中E1E2均參與結合,MXRA8的兩個結構域及鉸鏈區均與E1E2蛋白發生相互作用。隨后他們利用冷凍電鏡技術解析了人MXRA8CHIKV病毒樣顆粒的復合物結構,證明了MXRA8在病毒表面的結合模式跟晶體結構所觀察到的結合模式一致,并通過點突變及表面等離子共振(SPR)方法對結合關鍵氨基酸進行了驗證。該研究首次“看清”了基孔肯雅病毒和受體相互作用的分子模式,為一種新型的病毒-受體結合模式,同時糾正了過去對基孔肯雅病毒和受體相互作用的一些錯誤認識。

  MXRA8因其獨特的拓撲結構排布而與其他I型跨膜蛋白不同,其ND1結構域不是遠膜端結構域,而實為近膜端結構域。在復合物結構中,MXRA8D1結構域深深地插入到CHIKV三聚體刺突E蛋白的“峽谷”中,這就需要近膜端的頸部區有足夠的長度和柔性。

  通過一系列MXRA8莖部區截短體和病毒及細胞水平功能實驗發現,MXRA8中長達48個氨基酸的莖部區為病毒入侵所必需,其足夠長且具有柔性,被病毒利用作為受體入侵細胞。

  此項研究首次揭示了致關節炎甲病毒CHIKVMXRA8受體相互作用機制,證實了MXRA8是一種具有獨特的拓撲結構及結構域間組裝形式的新型Ig樣受體分子,并發現了此類致關節炎甲病毒和受體相互作用是一種新型的病毒-受體結合模式。這些研究結果為新型疫苗及廣譜中和抗體的研發提供了理論指導,為抗病毒藥物設計提供了新靶點。

  中科院北京生命科學研究院助理研究員宋豪及微生物所碩士生趙振楠、助理研究員柴彥為論文的并列第一作者,中科院院士高福和中科院天津工業生物技術研究所副研究員高峰為論文共同通訊作者。微生物所研究員齊建勛、施一、嚴景華及巴西FIOCRUZ基金會衛生技術發展中心主任Carlos Morel給予了大力支持。該研究得到科技部重點研發計劃、中科院戰略性先導科技專項項目、國家科技重大專項、國家自然科學基金以及中科院青促會、中國科協“青年人才托舉工程”等的經費支持。

  文章鏈接

 

1. 小鼠MXRA8的三維結構

 

2. CHIKV E與人MXRA8復合物結構

 

3. MXRA8莖部區對CHIKV入侵很關鍵

 

4. CHIKV利用MXRA8入侵細胞的分子機制模型




(責任編輯:葉瑞優)

附件:

專題推薦

相關新聞


© 1996 - 中國科學院 版權所有 京ICP備05002857號  京公網安備110402500047號  聯系我們

地址:北京市三里河路52號 郵編:100864

双彩网www.179012.com 975796.com | www.062697.com | www.262679.com | www.1126.me | www.330400.cc | www.325705.com | www.235435.com | www.938158.com | 博友彩www.842223.com | www.69870.com | 598742.com | www.xpj544.com | www.564480.com | www.25718.com | www.130.live | www.000333.cc | www.974500.com | 126482.com | www.4168.vip | www.070014.com | www.510494.com | www.035000.com | www.706111.cc | www.38my.com | www.043847.com | www.035649.com | www.457888.cc | 5863.biz | www.874720.com | www.444263.com | www.972841.com | 220312.com | 870773.com | www.lz07.com | www.211443.com | www.947949.com | 82385.com | 039078.com | www.634745.com | www.076615.com | www.810605.com | www.358717.com | www.332593.com | www.378140.com | www.305283.com | www.194068.com | 933708.com | 600万彩票www.187516.com | www.182827.com | www.687072.com | www.572257.com | 730906.com | www.8252.com | www.uo.com | www.891166.cc | 981461.com | www.739924.com | www.68em.com | www.081051.com | www.ch85.com | www.60117.com | www.008151.com | www.836517.com | www.044458.com | www.946.bid | www.968977.com | 864896.com | www.tw7.com | www.392782.com | 473384.com | www.86388.cc | www.661725.com | 23922.com | www.sn38.com | www.5396.org | www.76466.com | www.80289.com | www.402589.com | www.746902.com | www.60801.com | 967512.com | www.ck84.com | 908899.cc | www.71391.com | www.298662.com | www.446233.com | 268388.com | www.03572.com | www.989168.com | www.939301.com | 128413.com | www.981291.com | www.9eu.com | www.632.cm | www.270805.com | www.368229.com | www.568938.com | www.738262.com | www.808619.com | www.904585.com | 694433.com | www.xv06.com | www.595603.com | www.109516.com | www.268466.com | cai4411.com | 761346.com | www.296795.com | www.779353.com | www.ib95.com | www.325802.com | www.578469.com | www.732069.com | www.3872.wang | www.29859.cc | www.xpj811.com | www.315185.com | www.088988.cc | www.m34.tv | www.958047.com | 365065.cc | 974190.com | 787677.com | www.860909.com | www.772480.com | www.229129.com | www.805729.com | 281827.com | www.53336.cc | 548578.com | www.67uw.com | 大赢家彩票平台www.068631.com | www.1708.org | www.6367.xyz | 79sx.com | www.hw73.com | www.232291.com | www.834319.com | www.582837.com | www.212459.com | www.370502.com | 678277.com | 金利丰彩票www.630958.com | www.45bn.com | www.557688.com | www.590345.com | 834293.com | www.810633.com | www.118418.com | www.365420.cc | www.795512.com | 凤凰彩票www.88266x.com | www.82254.com | www.246331.com | www.776693.com | www.32zm.com | www.055398.com | www.968286.com | 975784.com | www.96kh.com | 798984.com | www.515751.com | www.38wc.com | www.58517.cc | www.471.cc | www.019810.com | www.957659.com | www.850776.com | 20444.com | 291169.com | 天天彩票www.206881.com | 99福彩www.729987.com | www.190035.com | www.288581.com | www.287896.com | www.10zm.com | www.45yj.com | js5023.com | 99彩票www.129029.com | www.98284.com | www.5520.win | www.vy63.com | www.99181.cc | 凤凰彩票www.77802h.com | www.72xm.com | www.34pr.com | www.60513.com | www.782778.com | www.401166.com | www.299615.com | www.287161.com | www.397708.com | www.3424.com | www.58157.cc | www.119300.com | 964484.com | www.ea24.com | www.9490.loan | www.56fb.com | www.617270.com | www.78238.com | www.940232.com | www.222942.com | www.348548.com | www.9858.net | 731498.com | www.841043.com | www.034500.com | www.36222.cc | www.006437.com | www.109505.com | www.996751.com | www.879805.com | www.pm35.com | www.111271.com | www.3oc.cc | www.233399.com | www.321272.com | www.924545.com | 951415.com | www.4388.net | www.138155.com | www.38362.cc | www.348969.com | 635702.com | 296679.com | www.770.cm | www.336892.com | www.756678.com | www.607955.com | 155164.com | www.844080.com | www.1054.cc | 358393.com | www.ms05.com | www.01206.cc | www.147789.cc | www.533751.com | 765309.com | www.567.gg | 128981.com | www.2211.live | www.75515.cc | www.073783.com | www.632935.com | www.03799.cc | www.002182.com | www.126825.com | www.2083.vip | www.197777.com | www.fq69.com | www.312698.com | www.44493.cc | www.6397.wang | www.260086.com | www.8734.loan | www.177977.com | www.367551.com | 552876.com | www.236553.com | www.05ce.com | www.948809.com | www.464482.com | www.566600.com | www.963354.com | www.819736.com | 107022.com | 587201.com | 773028.com | rv84.com | 盈发彩票www.99yfa.com | www.53182.cc | www.181183.com | www.016127.com | www.060822.com | www.59371.com | www.065308.com | www.230046.com | www.165613.com | www.rx04.com | 15574.com | www.065557.com | www.325529.com | 778579.com | 954715.com | 454477.com | TT彩票www.526093.com | 639205.com | www.qa4.cc | www.38xj.com | www.226019.com | www.631793.com | 世彩堂www.5854t.cc | www.cp6969.com | www.765535.com | www.656588.cc | www.14187.com | www.291278.com | 079358.com | 149571.com | www.155749.com | www.102115.com | www.580174.com | www.965647.com | www.505003.com | www.815981.com | 939725.com | 932080.com | 877694.com | www.283359.com | 博乐彩票www.35918w.com | www.034422.com | 120326.com | 653025.com | www.35125.cc | www.977201.com | www.488768.com | www.773532.com | www.942948.com | 739384.com | 420060.com | www.13bo.com | 680673.com | www.528598.com | www.277108.com | 497129.com | 吉利彩票www.80075h.com | www.97yx.com | www.74435.com | www.5622.com | 66677.com | 595851.com | www.788825.com | www.765731.com | www.273685.com | www.823378.com | www.975483.com | 853343.com | www.053928.com | www.620571.com | www.088693.com | www.260088.com | www.299072.com | www.358918.com | www.835688.com | www.957443.com | 81578.com | www.7038.biz | www.241569.com | www.770136.com | www.949120.com | www.899785.com | www.79ih.com | www.01666.com | www.696678.com | www.n05.net | www.903720.com | www.148681.com | www.808659.com | 569367.com | www.qz59.com | www.329059.com | www.885491.com | 696684.com | www.0429.me | www.594010.com | 795544.com | 28348.com | 877077.com | www.061023.com | www.049883.com | www.150504.com | www.09ni.com | www.0179.cc | www.973669.com | 627270.com | www.106739.com | www.807512.com | 025507.com | www.50ym.com | 997614.com | www.63dv.com | www.9qm.com | www.43390.com | www.496709.com | www.926507.com | www.756165.com | www.911113.com | www.36939.com | www.60jx.com | www.024534.com | www.4000.cm | www.76980.com | www.238558.com | www.746924.com | www.185154.com | www.2539.pw | www.039566.com | www.083489.com | www.197031.com | www.344163.com | www.bf10.cc | www.028162.com | www.440636.com | www.29828.com | www.151471.com | www.745277.com | www.655923.com | www.720992.com | www.654812.com | 026539.com | www.856872.com | 369684.com | 418896.com | 814233.com | 720735.com | www.129818.com | www.583630.com | www.iw44.com | www.pm24.com | www.1405.vip | www.031997.com | www.175870.com | www.316210.com | www.549633.com | www.871123.com | 205593.com | www.488521.com | www.661886.com | www.84rz.com | www.057179.com | www.068009.com | www.8097.loan | 800473.com | www.e17.bid | www.225199.com | www.365214.cc | www.030470.com | www.44490.cc | www.37285.cc | www.17725.com | www.8959.cn | www.267977.com | www.486528.com | www.108297.com | www.021356.com | www.473108.com | www.505290.com | www.580804.com | www.789089.com | www.851290.com | www.103714.com | www.529895.com | www.872591.com | 787586.com | www.403.press | 745172.com | 2566.com | 716985.com | www.87973.com | www.515721.com | www.453000.cc | www.485877.com | www.130710.com | www.036362.com | www.927580.com | 989847.com | www.80ud.com | www.91897.cc | www.915036.com | www.810098.com | www.691948.com | www.699050.com | www.900613.com | www.865571.com | www.824855.com | www.180460.com | www.567500.cc | 964517.com | www.4663.vip | www.397307.com | www.854338.com | www.dn52.com | www.792502.com | www.912219.com | 864462.com | www.yz55.com | www.5681.biz | www.2596.cn | www.cai353.com | www.yt43.com | www.35ig.com | www.411552.com | www.970643.com | www.438204.com | www.912358.com | 119142.com | www.456469.com | www.739038.com | www.861993.com | www.783788.com | www.981724.com | 038040.com | 豪彩VIPwww.30xw.com | www.cai2311.com | www.549.in | 933228.com | www.5388.mobi | www.2695.top | www.079087.com | www.cp982.com | www.84893.com | www.189758.com | www.700120.com | 961432.com | www.tf07.com | www.541290.cc | www.030721.com | www.492996.com | www.842404.com | www.925223.com | 872029.com | 222457.com | 38345.cc | 764977.com | 611246.com | www.vd83.com | www.67gz.com | www.4156.cm | www.0865.cc | www.75qd.com | www.138887.com | www.575185.com | www.670035.com | www.489109.com | www.621894.com | www.998336.cc | 117944.com | www.09td.com | www.718972.com | www.3163.vip | www.78495.com | www.359397.com | www.933989.com | www.779371.com | 971481.com | www.783856.com | 131807.com | www.392629.com | www.947929.com | www.968438.com | www.964443.com | www.855145.com | www.79oy.com | www.085808.com | www.69837.com | www.365701.com | www.655757.com | www.949315.com | www.891611.com | www.884034.com | www.sb88.cc | 幸运众彩www.108097.com | www.01094.com | www.122012.com | www.540034.com | www.862305.cc | www.5070.org | www.3367.vip | www.785672.com | www.xb1.com | 500彩票www.50054j.com | cp7116.com | www.85441.com | www.163367.com | www.528631.com | 388926.com | 易盈彩票www.7793x.com | www.85tx.com | www.569898.com | www.440119.com | www.335921.com | www.327130.com | www.196358.com | www.161651.com | www.090637.com | www.023335.com | www.3964.cm | www.049779.com | www.57913.com | www.4883.cn | www.1994.me | www.16fu.com | www.g69.xyz | www.fh88.com | www.xy3.com | www.498030.com | www.998233.com | 795219.com | www.198685.com | 692072.com | www.506088.com | www.195351.com | www.092063.com | www.629642.com | www.787798.com | 314788.com | 258443.com | www.131180.com | www.208770.com | www.015366.com | 7570.am | 906135.com | 972469.com | www.971270.com | 301129.com | www.e26.pw | www.6366.bid | www.0842.net | www.076508.com | www.489861.com | www.705977.com | 天天彩票www.54968f.com | 尊彩www.900140.com | 957435.com | www.06iy.com | www.3357.cn | 464087.com | 477710.com | www.153316.com | www.503779.com | www.590129.com | www.965893.com | www.111559.com | www.280014.com | www.176306.com | www.444883.com | www.004982.com | www.388764.com | 412328.com | www.667639.com | www.891053.com | www.818396.com | www.l22.shop | 648592.com | www.36691.cc | 506761.com--m1253.com | www.c86.biz | www.xg44.com | 8号彩票www.60108l.com | www.13de.com | 大赢家彩票平台www.976931.com | www.214508.com | www.251817.com | 029641.com | 743438.com | www.948705.com | www.782161.com | www.639610.com | www.608130.com | www.16486.com | www.216489.com | 518522.com | www.969209.com | www.822912.com | www.753348.com | 天天彩票www.2373j.com | 299823.com | 581428.com | 034983.com | www.133155.com | www.378719.com | www.672228.com | www.990506.com | www.003671.com | www.83946.com | www.196108.com | www.305516.com | www.539486.com | www.826977.com | 054058.com | 968140.com | 126497.com | www.213511.com | www.876803.com | www.991711.com | www.982928.com | 472543.com | www.759971.com | www.999141.com | www.v12.me | www.020542.com | 694118.com | www.891103.com | www.805306.com | www.912611.com | www.y80.biz | www.20jz.com | www.qh48.com | www.ci89.com | 707595.com | www.yb.com | 亿彩堂www.87668j.com | 博乐彩票www.35918a.com | 371994.com | www.cp3722.com | 90841.com | 金掌柜www.83993b.com | www.cp1155.com | www.cp5583.com | www.562455.com | www.42903.com | www.895039.com | 211369.com | 298854.com | 430830.com | www.275559.com | www.275687.com | www.263230.com | 222144.com | www.968347.com | www.866212.com | www.911519.com | www.596219.com | www.963521.com | 984586.com | www.ju81.cc | www.152244.com | www.540081.com | 947658.com | 17qe.com | 438231.com | www.225352.com | 4289.biz | www.113397.com | www.504896.com | www.503032.com | www.00wj.com | 673381.com | 500彩票www.3126l.com | 215351.com | www.p17.net | www.eb06.com | 110267.com | www.883430.com | www.456073.com | 982774.com | 974739.com | 365297.cc | www.199604.com | www.456480.com | www.qy16.com | www.169172.com | 497054.com | www.8854.loan | www.682721.com | www.686831.com | www.781523.com | www.nd5.com | www.773708.com | www.3074.biz | www.nd75.com | www.d26.hk | 312169.com | www.4xx.com | www.602876.com | www.633745.com | 191483.com | www.789391.com | 399949.com | www.732302.com | www.739454.com | www.725873.com | 365569.cc | 华彩www.71399p.com | www.g65.club | 99彩票www.577032.com | 121439.com | www.i53.club | www.24pv.com | www.7161.win | 155444.cc | www.pp95.com | www.879522.com | www.676765.com | www.670642.com | www.788647.com | www.216127.com | www.833150.com | 026039.com | www.88ht.com | www.7359.biz | www.ii5.com | www.95es.com | www.628262.com | www.01125.cc | www.090555.com | www.060653.com | 781413.com | www.83io.com | www.0000.top | www.079055.com | www.6465.cm | www.40140.com | www.68872.com | www.503601.com | www.444646.com | www.557879.com | www.626023.com | cai3377.com | 680886.com | www.559070.com | www.xz51.com | 240355.com | 735927.com | 579094.com | www.587948.com | www.357800.com | www.54581.com | www.091777.com | www.72wj.com | www.752018.com | www.679337.com | www.461898.com | www.422665.com | www.951716.com | www.532952.com | www.py5.com | www.lg69.com | www.541418.com | www.293097.com | 702308.com | www.203529.com | www.185499.com | 448001.com | 85fg.com | www.up16.com | www.43ir.com | www.zb97.com | www.5770.top | www.394.in | www.985390.com | www.911911.com | www.771115.com | www.674365.com | www.590751.com | www.4kt.cc | www.2446.com | www.2661.com | www.5777.in | www.83oo.com | www.04xq.com | www.207997.com | w1886.com | 好彩网www.889938.com | www.ng91.com | www.jz24.com | www.he.com | 943258.com | www.hj7.cc | www.jz55.cc | www.84wb.com | 660464.com | www.16dt.com | www.464657.com | www.751263.com | www.654826.com | www.729837.com | www.280772.com | 146558.com | 349535.com | 860402.com | 451989.com | www.522522.cc | www.615219.com | www.151559.com | www.137244.com | www.573113.com | www.494442.com | www.438805.com | www.944572.com | www.969002.com | www.884146.com | www.066.space | www.733.in | www.4776.cm | www.803789.cc | www.44vj.com | www.0307.in | www.898.me | 博友彩www.979683.com | 777692.com | 814629.com | 152678.com | www.861211.com | www.696110.com | www.548098.com | www.356024.com | www.231976.com | www.003256.com | www.039761.com | www.40445.com | www.6440.xyz | www.82ix.com | www.97yr.com | www.98rg.com | www.wb68.com | www.516227.com | www.468813.com | www.551.space | www.4429.vip | www.g78.org | 818合彩www.www.9889hc.com | www.968491.com | www.672757.com | www.782243.com | www.495751.com | www.17gk.com | 500彩票www.50026h.com | 488528.com | www.931411.com | 866015.com | 983305.com | 859067.com | www.319950.com | www.251058.com | 9275.biz | 72482.com | 120062.com | www.047323.com | www.332161.com | 973984.com | 19957.com | www.980961.com | www.669260.com | 784828.com | www.441089.com | 769408.com | 511901.com | 140386.com | www.684.in | www.jm04.com | 654238.com | 868644.com | www.910268.com | 190333.com | www.if54.com | www.tc03.cc | www.72qr.com | www.382.tv | 953425.com | www.407928.com | www.667220.com | www.446679.com | www.651408.com | www.914921.com | www.631819.com | 586489.com | 963460.com | 亿彩www.6832u.com | 趣彩网www.qucp0.com | 500彩票www.50054l.com | www.38232.com | 605988.com | www.78de.com | www.zr67.com | www.lz07.com | www.6945.vip | www.9020.vip | www.510.mobi | www.720.mobi | www.22321.com | 220524.com | 087805.com | www.169078.com | www.222385.com | www.0446.vip | www.022728.com | 342335.com | www.209.net | www.30860.com | www.044442.com | 0204.tv | www.973987.com | www.853707.com | www.780797.com | www.384056.com | www.209345.com | www.35127.cc | 彩票3www.808523.com | www.tp62.com | 005641.com | 613102.com | www.918143.com | www.739254.com | www.025921.com | www.35437.com | www.33417.com | www.60058.cc | www.98448.com | www.638.live | www.08977.com | www.551000.com | www.071433.com | www.178883.com | www.001508.com | www.107036.com | www.043816.com | www.003145.com | www.338426.com | www.393371.com | www.998580.com | www.325176.com | www.013997.com | www.836854.com | www.726.com | 978764.com | 960419.com | www.995959.cc | www.772733.com | www.639183.com | www.079163.com | www.829197.com | www.587072.com | 6687.com | 033702.com | www.446819.com | www.07nv.com | 490896.com | www.427986.com | www.485258.com | www.187589.com | www.44302.com | www.21wj.com | 福彩网www.5522r.cc | www.fm24.com | www.cai7266.com | 73929.com | www.237263.com | www.051926.com | www.8886.me | www.959186.com | www.799822.com | www.665764.com | www.526731.com | www.301988.com | www.58368.com | www.9930.top | www.019130.com | www.110594.com | www.791396.com | www.628755.com | www.213246.com | www.55275.com | www.009520.com | www.3199.cc | www.cp1516.com | www.066259.com | www.69094.com | www.14040.com | www.1844.net | www.402638.com | 8cr.cc | www.894919.com | www.518781.com | www.313530.com | www.397673.com | www.672030.com | www.69ii.com | www.961558.com | www.773008.com | 841337.com | cz40.com | 376086.com | 022363.com | www.838501.com | www.09zk.com | www.17622.com | www.29897.com | www.e82.net | www.8036.cn | www.cp372.com | 659286.com | www.903633.com | www.622773.com | www.681956.com | www.892302.com | www.641314.com | www.226493.com | www.097195.com | www.37656.cc | www.165.win | www.974742.com | www.772836.com | www.183904.com | www.373791.com | www.442811.com | www.290667.com | www.232177.com | www.67988.com | www.959451.com | www.696320.com | www.503231.com | www.165621.com | www.231308.com | www.231608.com | www.204068.com | www.003564.com | www.7867.top | www.536.name | www.15nf.com | www.fv54.com | www.568.tv | www.17is.com | 99彩票www.29699.com | www.873820.com | 870267.com | 015784.com | www.506561.com | www.486693.com | www.678352.com | www.984347.com | www.688561.com | www.co50.com | www.su64.com | www.39du.com | www.cai911.com | www.246535.com | www.395915.com | www.147988.com | 220108.com | www.973747.com | www.qa3.com | 021758.com | www.801139.com | www.573234.com | www.192713.com | www.4824.cc | www.cp559.com | 693007.com | 599346.com | www.862323.com | www.5550.in | www.49sh.com | www.cai2227.com | 彩神购彩www.csgc7.com | 949675.com | 846816.com | www.632357.com | 亚投彩票www.ya515.com | www.958201.com | www.356450.com | www.299726.com | www.556981.com | www.215383.com | www.t06.cc | www.334677.com | 028470.com | www.77220.cc | www.24374.com | www.5544.biz | www.uf9.cc | 049351.cc | www.781549.com | www.843844.com | www.062386.com | www.023618.com | www.037186.com | www.147698.com | www.143573.com | 顶尖彩票www.168212.com | uq36.com | www.933385.com | 311123.com | www.871101.com | www.036235.com | www.5252.la | www.138506.com | www.123550.com | www.46552.com | www.4415.vip | www.bb83.com | 126630.com | 354009.com | www.915850.com | www.011007.com | www.418055.com | www.250042.com | www.236822.com | www.41768.com | www.00951.com | www.815788.com | www.744325.com | www.460099.com | www.668199.com | www.948442.com | www.713691.com | www.441189.com | www.wp32.com | 954849.com | www.ue94.com | 436755.com | www.131234.cc | www.83te.com | 成功彩票www.le33.cc | 277743.com | www.057178.com | www.067943.com | www.2029.xyz | www.75657.com | www.14895.com | 910474.com | www.958872.com | www.812108.com | 858267.com | www.50933.com | www.222474.com | www.040537.com | www.7147.tv | www.78178.cc | www.356096.com | www.164132.com | www.007396.com | www.340991.com | www.381747.com | www.152101.com | www.531852.com | 110791.com | 558513.com | www.570036.com | 967366.com | www.824234.com | www.739938.com | 947381.com | www.124.cx | www.982431.com | www.539854.com | www.321768.com | 898168.com | www.633408.com | www.954802.com | 004035.com | www.771145.com | www.618331.com | www.77uz.com | 570959.com | 78035.com | www.948740.com | www.700936.com | www.487021.com | www.967178.com | www.851296.com | www.971315.com | www.709138.com | www.507233.com | www.968223.com | www.589875.com | www.71ih.com | vo57.com--2979.com | www.655377.com | www.990646.com | www.401544.com | www.068.net | 133008.com | www.981228.com | www.506644.com | www.35437.com | 好运来www.998855k.com | www.870133.com | www.354664.com | www.89263.com | www.gc06.com | www.4964.cc | www.bi1.cc | www.30mc.com | www.211266.com | www.7632.cn | k3k3彩票www.602952.com | 388690.com | www.917358.com | www.678129.com | www.236938.com | www.039507.com | www.114866.com | www.07iv.com | www.731094.com | www.220084.com | www.869890.com | www.9531.org | www.zj3.com | 028184.com | www.774606.com | www.800198.com | www.ih3.cc | 001152.com | www.646539.com | www.566522.com | www.010482.com | www.zv40.com | www.32799.com | www.759098.com | www.294244.com | www.04180.com | www.296.cm | www.4413.cn | www.5829.site | www.06ey.com | 791085.com | www.976053.com | www.520727.com | www.008264.com | www.16oc.com | kw30.com--kw30.com | www.737256.com | www.111438.com | www.3588.vip | www.521475.com | www.166145.com | www.8273.xyz | www.85256.com | www.9197.online | www.00cf.com | 595236.com | 727242.com | www.852863.com | 802931.com | www.947149.com | www.554107.com | www.371010.com | www.932880.com | www.do4.cc | www.931619.com | www.383088.com | www.008749.com | www.840078.com | www.544111.com | www.209097.com | 498325.com | www.772846.com | www.179590.com | 大赢家彩票www.599894.com | www.78ce.com | ah39.com | www.212820.com | www.62913.cc | www.089045.com | www.8845.cc | www.6124.vip | 289217.com | www.850608.com | www.668718.com | www.859083.com | www.531670.com | www.065098.com | www.p83.com | www.63ax.com | 57dg.com | 89456.com--444409.com | www.557176.com | www.039900.com | 919339.com | www.404081.com | www.132462.com | www.93735.com | www.7468.cn | 彩多多www.78949k.com | 846517.com | www.823550.com | www.445220.com | www.024624.com | www.721.live | www.39659.cc | www.29hm.com | 乐趣彩www.lqc2.com | www.144708.com | www.26212.cc | www.qd14.com | www.hj19.com | www.45ff.com | www.41011.com | www.244014.com | www.43hl.com | www.6206.vip | www.cp7998.com | www.028760.com | www.10uw.com | 162406.com | 988794.com | www.15nx.com | 518200.com | www.954748.com | www.229816.com | www.190071.com | www.220536.com | 659634.com | www.989598.com | 964943.com | 246379.com | www.883826.com | www.867682.com | www.731811.com | www.869257.com | www.760270.com | www.208187.com | www.8900.vip | 99796.com | www.929094.com | www.523733.com | www.103165.com | www.1237.cn | www.43bz.com | 126584.com | www.918218.com | www.881694.com | www.875262.com | www.771803.com | www.631123.com | www.654138.com | www.455001.com | www.091673.com | www.517.tv | www.zz43.cc | 897611.com | www.965638.com | www.29lq.com | www.f86.pw | 584391.com | www.987606.com | www.620876.com | www.739214.com | www.989175.com | www.502377.com | www.56925.cc | www.458021.com | www.211066.com | www.813095.com | www.cp0374.com | www.943800.com | www.505945.com | www.568200.com | www.75117.cc | www.473551.com | www.118566.com | www.58727.cc | www.87od.com | 996639.com | 982546.com | www.715297.com | www.213248.com | www.007458.com | www.4756.com | 598221.com | www.17193.cc | www.858431.com | www.520436.com | 578251.com | 670496.com | www.777336.com | www.403643.com | www.507278.com | rd57.com | www.119757.com | www.55552.cc | www.82837.cc | www.17oy.com | www.220035.com | www.6054.vip | www.er00.com | 758778.cc | www.941162.com | www.517025.com | www.397838.com | 801674.com | www.975858.com | www.607693.com | 137608.com | www.563392.com | 858436.com | www.553266.com | www.081910.com | www.25rb.com | www.684.xyz | www.oc69.com | www.755871.com | www.624302.com | www.636722.com | www.829459.com | www.907976.com | www.413311.com | www.gg1.cc | www.hu57.com | www.853046.com | www.759576.com | www.248978.com | www.337660.com | www.46au.com | 040851.com | 608561.com | www.067124.com | www.373667.com | www.713167.com | www.543997.com | www.50595.com | www.qc82.com | www.984347.com | www.320488.com | www.9432.xyz | 3265.biz | www.720338.com | 443653.com | www.61kg.com | 386411.com | www.948017.com | www.465939.com | www.165448.com | www.019664.com | www.159327.com | www.434986.com | www.099576.com | www.780816.com | www.955155.com | www.589610.com | www.486999.com | www.562187.com | 422514.com | 4355.biz | www.94993.com | www.191529.com | www.103845.com | www.1378.mobi | www.73ho.com | www.79ei.com | www.398691.com | www.69654.com | 54bu.com | www.918473.com | www.020703.com | 361817.com | www.788398.com | www.209251.com | www.81332.cc | www.737038.com | 577977.com--331777.com | www.924147.com | www.385102.com | www.975941.com | www.590075.com | 606215.com | www.635626.com | www.012222.com | www.hg55.com | 712669.com | www.959267.com | www.582721.com | www.077288.com | www.003057.com | www.cw92.com | www.369354.com | www.325448.com | www.16ln.com | 848198.com | 982354.com | 778034.com | www.860145.com | www.616491.com | 038849.com | www.02940.cc | www.288257.com | www.068123.com | www.057893.com | www.0137.top | www.230220.com | www.813656.com | www.491812.com | www.117534.com | www.21591.cc | www.463444.com | www.003801.com | www.71zp.com | www.32977.com | www.84io.com | 026836.com | www.719135.com | www.8690.cc | www.175951.com | www.853968.com | www.466546.com | www.qs55.com | www.157654.com | www.093977.com | www.49tm.cc | www.0547.win | aa98.cc | www.778773.com | www.39737.cc | www.3515.red | 948418.com | www.118370.com | www.3227.shop | 众彩网www.180435.com | www.923728.com | 067780.com | www.799650.com | www.947066.com | www.685556.com | www.777496.com | www.386207.com | www.595809.com | www.854811.com | www.204002.com | www.12460.com | www.0315.xyz | www.gv59.com | www.673447.com | 756466.com | 796764.com | www.138520.com | www.754068.com | www.067608.com | www.822644.com | www.490636.com | www.nb16.com | www.st2.com | www.244887.com | www.957230.com | 295949.com | www.781851.com | www.732300.com | 753106.com | www.800104.com | www.570719.com | www.57761.cc | www.qx63.com | www.407100.com | www.326038.com | 788399.cc | www.485131.com | 50485.com | www.751272.com | www.617811.com | www.4737.com | 9761.biz | www.599833.cc | www.839583.com | 汇彩网www.hc5859.com | 260748.com | www.087550.com | www.567129.com | www.142758.com | www.773830.com | 383481.com | 026494.com | www.822530.com | 545537.com | www.506351.com | www.165186.com | www.077345.com | www.9vb.com | 358147.com | 461489.com | www.188912.com | 894857.com | www.932840.com | www.664111.com | www.345086.com | www.29931.cc | www.163955.com | 发彩www.50732y.com | www.601702.com | www.8325.biz | 087466.com | 竞彩网www.8jcw.com | 网易彩票www.501209.com | www.925229.com | www.485866.com | www.77424.com | www.g35.org | www.9166.top | 吉利彩票www.80065n.com | www.5876.vip | 大赢家彩票平台www.810926.com | 544185.com | www.301000.com | www.0635.top | www.7452.cn | 072089.com | www.737368.com | www.179691.com | www.220503.com | www.01907.com | 彩票坊www.022qh.cc | www.969078.com | www.161758.com | 258817.com | 1887.pw | www.q15.net | 815631.com | www.10321.cc | 环球彩票www.81455.com | 610289.com | www.963653.com | www.285227.com | www.45702.com | www.mt21.com | 527936.com | www.969763.com | www.329356.com | www.ak20.com | www.135848.com | www.6338.cm | www.151039.com | www.27vd.com | 693036.com | www.541382.com | www.036850.com | www.pg16.com | bwin6688.com | www.11oq.com | 240250.com | www.629192.com | www.099987.com | www.gy31.com | www.52787.cc | www.sv07.com | www.23371.cc | www.013210.com | www.up61.com | www.2599.cc | 055064.com | www.680899.com | www.135991.com | www.1lf.cc | www.419200.cc | www.53852.cc | 935075.com | www.988874.com | 385406.com | www.yv73.com | www.974371.com | www.997564.com | www.562591.com | www.866428.com | www.826099.com | www.594770.com | www.4544.vip | 035171.com | www.756848.com | www.598428.com | www.90460.com | www.85rw.com | 602959.com | www.749053.com | www.op21.com | 527921.com | 714571.com | www.717845.com | www.433636.com | www.2034.vip | 07cu.com | www.72of.com | 739047.com | 031939.com | www.76399.com | www.293289.com | www.47uf.com | www.4951.org | 805080.com | 497985.com | www.790924.com | www.067914.com | www.011092.com | www.252708.com | www.581700.com | www.37ps.com | www.367003.com | www.585485.com | www.310.net | www.157764.com | www.98qs.com | www.1080.mobi | 607495.com | 006484.com | 279898.com | www.559316.com | www.138060.com | www.131415.com | www.9071.vip | 434777.com | www.822530.com | www.258053.com | www.20718.com | www.697.date | www.1410.biz | www.482365.com | www.08087.cc | www.mi7.cc | www.874495.com | www.708936.com | www.263958.com | www.3434.cm | www.bg02.cc | 9283.net | www.848483.com | www.130519.com | www.60qc.com | 327115.com | 028713.com | www.669892.com | www.mf77.com | www.956801.com | www.947022.com | www.542059.com | www.738208.com | www.082257.com | www.x85.org | www.509684.com | www.676330.com | www.152298.com | www.cw66.com | www.955802.com | www.774416.com | www.837158.com | www.mb39.com | www.7676.vip | 427077.com | www.485709.com | www.964990.com | www.214408.com | www.843943.com | 107220.com | www.730629.com | www.039048.com | www.018932.com | www.696951.com | www.69607.com | 762532.com | www.881986.com | www.438917.com | www.4410.vip | www.543563.com | www.nm88.com | 830325.com | www.864200.com | www.922290.com | www.591136.com | www.4336.cc | 9916.me | www.987520.com | www.257092.com | www.6251.biz | 949954.com | www.876710.com | www.kw70.com | www.998797.com | www.437380.com | www.329223.com | 543350.com | www.366716.com | www.72ye.com | 500彩票www.3126b.com | 744064.com | www.125689.com | 936608.com | www.595290.com | www.016304.com | 天天彩票www.106316.com | www.575339.com | 029537.com | www.0243.in | www.088082.com | www.25pz.cc | www.873575.com | www.888999.com | www.255965.com | www.6429.org | www.264342.com | www.27791.cc | www.1ky.com | www.228885.com | www.32si.com | www.zo57.com | www.414178.com | www.854406.com | www.867757.com | 982797.com | www.87653.cc | 338453.com | www.bv33.com | 313580.com | www.53dr.com | 302325.com | www.592712.com | www.19832.cc | www.rf75.com | www.1787.tv | www.609332.com | www.5317.online | 虹彩集团www.hcjt2.com | www.762097.com | www.274744.com | www.13983.com | 288319.com | www.860496.com | www.299909.com | www.25lz.com | 博乐彩票www.684335.com | www.812664.com | www.764789.com | www.166852.com | 702875.com | www.216616.com | www.2464.win | www.290977.com | www.49do.com | www.976765.com | 358438.com | www.385449.com | 88948.com | www.441443.com | www.7095.biz | www.rp31.com | cai4545.com | www.637583.com | www.317369.com | www.47354.com | www.cx77.com | www.977164.com | www.282685.com | www.99159.cc | www.6869.biz | 136801.com | www.545679.com | www.32833.cc | www.450821.com | www.8079.vip | 030922.com | www.974547.com | www.387595.com | 445654.com | www.845813.com | www.91553.cc | 奔驰彩票www.750966.com | 546103.com | www.321765.com | www.3053.vip | 842883.com | www.0255.cm | www.861.win | 156567.com | www.48388.com | www.50dj.cc | 6362.biz | www.770198.com | www.903257.com | www.168697.com | www.7to.com | www.870642.com | www.811684.com | www.2236.cn | www.64fh.com | www.057818.com | www.cai5266.com | www.701126.com | www.769569.com | www.93696.cc | www.645107.com | 734306.com | www.728018.com | www.58672.com | 255089.com | 807607.com | 917501.com | www.649446.com | www.19662.cc | www.35291.cc | ou85.com | www.134843.com | www.637.cc | www.258273.com | www.lr03.com | www.901517.com | www.036279.com | 598571.com | www.887539.com | www.223345.com | www.dq36.com | www.972587.com | 036009.com | www.986782.com | www.350210.com | www.74847.com | www.mh9.com | www.777842.com | 913071.com | www.609603.com | www.m18.cn | www.956275.com | www.998664.com | www.233414.com | www.15455.cc | www.1969.net | 604867.com | www.623217.com | www.629885.com | www.39681.cc | 954467.com | www.474801.com | www.223196.com | www.84sj.com | www.998934.com | www.978123.com | www.189240.com | www.169164.com | www.ry40.com | 456322.com | www.12iz.com | www.864660.com | www.465922.com | www.457062.com | www.1546.vip | 130259.com | www.700966.com | www.588999.com | www.912535.com | www.929588.com | www.104544.com | www.cv.com | www.166237.com | www.09lk.com | www.73801.com | www.cai4242.com | 266901.com | www.770268.com | 303659.com | www.829759.com | www.132749.com | www.36jy.com | 72749.com | www.cm55.com | www.106980.com | www.335207.com | www.64uo.com | www.734178.com | 750954.com | www.0175.xyz | www.c14.space | www.863125.com | www.582376.com | www.637472.cc | www.952490.com | www.749029.com | www.181202.com | www.1519.vip | 814601.com | www.553982.com | www.1040.vip | www.279058.com | www.95992.cc | 654275.com | www.317348.com | www.867182.com | www.cai7266.com | np47.com | 980547.com | www.584841.com | www.318084.com | www.026114.com | ys58.com | www.579541.com | www.7370.loan | 713520.com | www.536525.com | www.7346.vip | 918彩票网www.918jo.com | www.689281.com | 天天彩票www.54968f.com | www.699718.com | 135111.cc | www.588853.com | www.67843.com | www.69235.com | www.60um.com | www.vx37.com | www.563317.com | 96qs.com | www.778089.com | www.974897.com | www.886952.com | www.k17.biz | 024012.com | www.dr84.com | 129754.com | www.571088.com | 975941.com | www.702707.com | www.043946.com | 853824.com | www.504778.com | www.9200.cm | www.060044.cc | www.8yy.cc | www.43788.com | www.mp5.com | www.27kg.com | 975745.com | www.586855.com | 947848.com | www.846291.com | 240638.com | www.757468.com | www.968290.com | www.457499.com | 501624.com | www.775445.com | www.170457.com | www.286860.com | 738939.com | www.620305.com | www.9138.net | www.984883.com | www.151212.com | 9亿彩票www.857t.com | www.611573.com | www.7638.cn | 400604.com | www.320328.com | www.xy94.com | www.782349.com | www.000863.com | cai3366.com | www.730157.com | www.59297.com | 266902.com | www.340398.com | www.35eh.com | www.874155.com | www.088194.com | 中国福利彩票www.08588x.com | www.972576.com | www.351238.com | www.xh18.com | www.665216.com | www.99474.com | www.aj47.com | www.689888.com | www.37635.cc | 722852.com | www.289363.com | 易彩网www.12455y.com | www.187296.com | www.69136.com | 797672.com | 799605.com | www.448180.com | 988338.com | www.603920.com | www.981852.com | www.230711.com | www.825916.com | www.795759.com | www.788564.com | www.40562.com | 257977.com | www.282629.com | www.010234.com | 358759.com | www.320307.com | www.385027.com | www.351.tv | 848245.com | www.775656.com | www.794334.com | www.820949.com | www.516567.com | www.20rf.com | www.774064.com | www.170990.com | www.63zo.com | www.963072.com | www.168998.com | www.7782.org | 4159.biz | www.809980.com | www.748893.com | www.843844.com | www.591164.com | www.4689.me | www.27752.com | 967148.com | www.530589.com | www.189742.com | www.32go.com | TT彩票www.356036.com | www.332380.com | www.4559.cn | 74055.com | www.390091.com | www.19ax.com | www.737308.com | www.053820.com | 218678.com | www.11qc.com | 970146.com | www.692176.com | 023784.com | www.zb47.com | www.042644.com | www.bu2.cc | www.635569.com | www.453234.com | www.4145.cc | www.996047.com | www.215860.com | www.07qv.com | www.878674.com | www.724019.com | www.89798.cc | www.298569.com | www.57gn.com | www.sn29.com | www.756521.com | www.237393.com | 网易彩票www.097wy.com | www.665958.com | www.4628.net | 265016.com | www.338574.com | www.22959.cc | www.96hk.cc | www.986034.com | www.143856.com | www.659400.com | www.171108.com | www.0233.me | he48.com | www.715834.com | www.006917.com | 303366.com | www.967495.com | www.483815.com | www.036543.com | 627026.com | 589055.com | www.308598.com | 237349.com | www.937773.com | www.413100.cc | www.128928.com | www.cp1103.com | www.378442.com | www.82836.cc | 彩虹8www.ch8988.com | www.759862.com | www.001785.com | www.eu41.com | www.755511.com | E彩堂www.43818e.com | 281219.com | www.488633.com | www.04137.com | 572891.com | www.961109.com | www.98100.com | www.322418.com | www.cp124.cc | www.602121.com | www.003957.com | www.220668.com | www.100230.com | www.gb24.com | www.345444.cc | www.286644.com | www.412087.com | www.92kj.cc | www.89223.cc | 4111vip.com | www.124365.com | 265418.com | www.556580.com | 000724.cc | www.636032.com | www.93978.cc | pe3.cc | www.569000.com | www.56686.com | www.ek08.com | www.710895.com | www.790636.com | www.590558.com | www.3549.cm | www.473618.com | www.95ue.com | 570081.com | www.432131.com | www.778746.com | www.086377.com | www.1896.hk | www.cai888.com | www.659327.com | www.722405.com | www.292390.com | www.43pi.com | www.fj79.com | 005424.com | www.ji77.com | www.866624.com | www.522583.com | www.34687.com | www.938311.com | 500彩票www.549377.com | www.923292.com | www.200988.cc | www.60md.com | www.905027.com | www.101325.com | 购彩堂www.680352.com | www.1591.vip | 4930